Study on mitotic and meiotic homologous recombination in plant and cultured cells of rice

水稻植株和培养细胞有丝分裂和减数分裂同源重组的研究

基本信息

批准号：
15580001
负责人：
NIIZEKI Minoru
金额：
$ 2.37万
依托单位：
Hirosaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580001/
关键词：
homologous recombination RiLIM15 Alternative splicing Meiosis southern blot Exon Intron gene family 相同染色体組換え遺伝子 mRNA cDNA ソマクローナル変異培養細胞 OsRAD51 相同染色体組換えイネゲノムモチ・ウルチ分離比

项目摘要

1.Analysis of occurrence of MCO in cultured cell of rice : Calli were induced from F_1 seed (Wxwx) of glutinous rice (wxwx) as a seed parent and non-glutious rice(WxWx) as a pollen parent. After regeneration of rice plamts, it was found a dominant homologous plant(WxWx) which might be segregated by the MCO.2.Analysis of structure and expression pattern of RiLIM15 in rice : In order to clarify the relationships between occurrence of MCO and expression of RiLIM15 gene in cultured cells and panicles investigated on the aspects of structure and expression of RiLIM15. Two RiLIM15 homologous RiLIM15A and RiLIM15B were isolated. In exon region, the DNA sequences for two genes were highly homologous but not intron regions. The amino acid sequences these two genes were highly homologous, sugggesting that they may be nearly equivalent in function. Analysis of cDNA sequences that eight patterns of deletions of DNA sequences in RiLIM15 cDNA clones derived from both meiotic and cultured cells. They may be resulted from alternative splicing. The putative mRNA with these deletion may translate different functions of proteins.3.Analysis of RAD51 in rice : RT-PCR was performed using total RNA extracted from the cultured cells, young meiotic panicles, mature leaves and X-ray treated seedlings. Expression of the RAD51 gene was found to occur in cultured cells, young meiotic panicles and X-ray treated seedlings, but not mature leaves. The gene possibly participates in DNA repariring of MCO.4.Application to substantial plant breeding : In nine generation of progenies regenerated from calli of cv.Akihikari it was observed that a line is 3-4 early heading date and almost the same number of panicle as the parent cultivar and significantly higher thousand kernel weight than the parent cultivar.

1.水稻培养细胞中MCO发生情况分析：以糯米(wxwx)的F_1种子(Wxwx)为种子亲本，粳米(WxWx)为花粉亲本诱导愈伤组织。水稻再生后，发现了一个可能被MCO分离的显性同源植物（WxWx）。2.水稻RiLIM15的结构和表达模式分析：为了阐明MCO的发生与MCO表达之间的关系。对培养细胞和圆锥花序中RiLIM15基因的结构和表达进行了研究。分离出两个RiLIM15同源物RiLIM15A和RiLIM15B。在外显子区域，两个基因的DNA序列高度同源，但内含子区域则不同。这两个基因的氨基酸序列高度同源，表明它们在功能上可能几乎相同。对 cDNA 序列进行分析，发现来自减数分裂细胞和培养细胞的 RiLIM15 cDNA 克隆中 DNA 序列缺失的八种模式。它们可能是由选择性剪接产生的。具有这些缺失的推定mRNA可能翻译蛋白质的不同功能。3.水稻中RAD51的分析：使用从培养细胞、减数分裂幼穗、成熟叶片和X射线处理的幼苗中提取的总RNA进行RT-PCR。研究发现，RAD51 基因的表达出现在培养细胞、年轻的减数分裂圆锥花序和 X 射线处理过的幼苗中，但不出现在成熟的叶子中。该基因可能参与MCO的DNA修复。 4.在植物实质育种中的应用：在cv.Akihikari愈伤组织再生的9代后代中，观察到一个品系的抽穗期早3-4个，穗数几乎相同。与亲本品种相比，千粒重显着高于亲本品种。